Skip to main content

Cell Culture Assays

  • 427 Accesses

Part of the Springer Protocols Handbooks book series (SPH)


Cell culture assays play an important role in drug research along with animal tests with the principle of ‘3Rs’—Reduction, Refinement and Replacement—due to its quick and easy way of drug response analysis, as well as high throughput in assays, simplicity and cost-effectiveness. This chapter explains various procedures, cell viability, cytotoxicity and inflammatory response, for the evaluation of cytotoxicity of novel compounds or extracts. The protocols like propidium iodide used cell cycle assay and apoptosis assay, Alamar Blue method in Caco-2 cells, cell viability test in Caco-2 cells using lipopolysaccharide (LPS) and cytotoxicity against RAW 264.7 for the determination of anti-inflammatory activity—together with the assays for nitric oxide (NO) determination and scratch assay for testing the proliferation of fibroblast cells and inflammatory response in AGS cells model—elucidate the detailed and standardized protocols for the in vitro assays with cell lines.

Key words

  • Alamar Blue
  • NO
  • LPS
  • AGS
  • Scratch assay
  • Fibroblast cells
  • Propidium iodide
  • Annexin V

This is a preview of subscription content, access via your institution.

Buying options

USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-0716-1233-0_2
  • Chapter length: 17 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-0716-1233-0
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   139.99
Price excludes VAT (USA)
Hardcover Book
USD   179.99
Price excludes VAT (USA)

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more


  1. Woo CSJ, Lau JSH, El-Nezami H (2012) Herbal medicine: toxicity and recent trends in assessing their potential toxic effects. In: Advances in botanical research, vol 62. Academic Press, Cambridge, MA, pp 365–384

    Google Scholar 

  2. Wu MH, Huang SB, Lee GB (2010) Microfluidic cell culture systems for drug research. Lab Chip 10(8):939–956

    CAS  CrossRef  PubMed  Google Scholar 

  3. Rumjanek VM, Trindade GS, Wagner-Souza K, Meletti-de-Oliveira MC, Marques-Santos LF, Maia RC, Capella MA (2001) Multidrug resistance in tumour cells: characterisation of the multidrug resistant cell line K562-Lucena 1. An Acad Bras Cienc 73(1):57–69

    CAS  CrossRef  PubMed  Google Scholar 

  4. Joe AK, Liu H, Suzui M, Vural ME, Xiao D, Weinstein IB (2002) Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines. Clin Cancer Res 8(3):893–903

    CAS  PubMed  Google Scholar 

  5. Nakayama GR, Caton MC, Nova MP, Parandoosh Z (1997) Assessment of the Alamar Blue assay for cellular growth and viability in vitro. J Immunol Methods 204(2):205–208

    CAS  CrossRef  PubMed  Google Scholar 

  6. McGaw LJ, Elgorashi EE, Eloff JN (2014) Cytotoxicity of African medicinal plants against normal animal and human cells. In: Toxicological survey of African medicinal plants. Elsevier, Amsterdam, pp 181–233

    CrossRef  Google Scholar 

  7. Xu B, Li YL, Xu M, Yu CC, Lian MQ, Tang ZY, Li CX, Lin Y (2017) Geniposide ameliorates TNBS-induced experimental colitis in rats via reducing inflammatory cytokine release and restoring impaired intestinal barrier function. Acta Pharmacol Sin 38(5):688–698

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  8. Joo T, Sowndhararajan K, Hong S, Lee J, Park SY, Kim S, Jhoo JW (2014) Inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells by stem bark of Ulmus pumila L. Saudi J Biol Sci 21(5):427–435

    CrossRef  PubMed  PubMed Central  Google Scholar 

  9. Dimitrios B (2006) Sources of natural phenolic antioxidants. Trends in food science & technology 17(9):505–512

    Google Scholar 

  10. Gerlier D, Thomasset N (1986) Use of MTT colorimetric assay to measure cell activation. J Immunol Methods 94(1–2):57–63

    CAS  CrossRef  PubMed  Google Scholar 

  11. Plumb JA (2004) Cell sensitivity assays: the MTT assay. In: Cancer cell culture. Humana, Totowa, NJ, pp 165–169

    Google Scholar 

  12. de Freitas Figueiredo F, Cechinel Filho V, Damazo AS, Arunachalam K, Colodel EM, Ribeiro M et al (2020) Sorocea guilleminiana Gaudich.: wound healing activity, action mechanisms, and chemical characterization of the leaf infusion. J Ethnopharmacol 248:112307

    CrossRef  Google Scholar 

  13. Suffness MP, Pezzuto JM (1990) Assays related to cancer drug discovery. In: Hostettman K (ed) Methods in plant biochemistry: assays for bioactivity. Academic, Cambridge, MA, pp 71–133

    Google Scholar 

  14. Khlifi D, Sghaier RM, Amouri S, Laouini D, Hamdi M, Bouajila J (2013) Composition and anti-oxidant, anti-cancer and anti-inflammatory activities of Artemisia herba-alba, Ruta chalpensis L. and Peganum harmala L. Food Chem Toxicol 55:202–208

    CAS  CrossRef  PubMed  Google Scholar 

  15. Park E, Kum S, Wang C, Park SY, Kim BS, Schuller-Levis G (2005) Anti-inflammatory activity of herbal medicines: inhibition of nitric oxide production and tumor necrosis factor-α secretion in an activated macrophage-like cell line. Am J Chin Med 33(03):415–424

    CrossRef  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations


Rights and permissions

Reprints and Permissions

Copyright information

© 2021 The Author(s), under exclusive license to Springer Science+Business Media, LLC , part of Springer Nature

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Arunachalam, K., Sasidharan, S.P. (2021). Cell Culture Assays. In: Bioassays in Experimental and Preclinical Pharmacology. Springer Protocols Handbooks. Humana, New York, NY.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1232-3

  • Online ISBN: 978-1-0716-1233-0

  • eBook Packages: Springer Protocols